Objective To explore the effect of hypoxia-ischemia(HI)on expression of second mitochondria-derived activator of caspase proteinc(Smac)in brain tissue of neonatal rats.Methods All neonatal rats were divided randomly into HI group,pseudo-operation group,and normal control group.The animal models of HIE were made.At 24,48,72,96 h after HI,Western blot was used to detect the expression of Smac protein in hippocampus.The relative activity of Caspase-3 in hippocampus tissue was determined by colorimetric assay.Results In HI group,at 24,48,72,96 h after HI,the expression levels of Smac protein in hippocampus significantly increased compared with that of pseudo-operation group and normal control group(Pa

OBJECTIVETo develop a method for determining epichlorohydrin in the workplace air by gas chromatograph-electron capture detector (GC-ECD).

METHODSEpichlorohydrin in the workplace air was collected by activated charcoal tubes, desorbed using acetone, and analyzed by GC-ECD.

RESULTSA good linearity was obtained in the range of 1.0-50 µg/mL (r=0.999 7). The detection limit was 0.012 µg/ml, while the recovery rate was 88.1% and relative standard deviation ranged from 1.11% to 3.57%. The samples could be stored for seven days at room temperature.

CONCLUSIONThis method effectively eliminates the interferences of alkanes on determination of epichlorohydrin and improves the sensitivity by 1 to 2 orders of magnitude, which can solve the problem of detection limit above standard in GBZ/T 160.58-2004.

Air Pollutants, Occupational ; analysis ; Charcoal ; Chromatography, Gas ; Epichlorohydrin ; analysis ; Workplace

Objective: Changes in the ?2 integrin of adhesion molecules between cells are closely associated with the invasion and migration of tumor cells.This study aimed at the effect of anti-?2 integrin on the invasion and migration of osteosarcoma MG63 cells. Methods: The osteosarcoma MG63 cell line was cultured in the DMEM medium.The effect of the anti-?2 integrin monoclonal antibody on the migration and invasion of tumor cells were measured by scratch assay and Transwell assay.The migration and invasion cells were stained by Crystal violet staining and counted under the hundredfold microscope.Results: Compared with the control group,the migration and invasion abilities of the MG63 cells were significantly decreased in the anti-?2 treatment groups.Conclusion: Anti-?2 integrin may inhibit the migration and invasion of osteosarcoma cells.

Objective To explore the roles and possible mechanism of calcium-sensing receptor(CaSR) in cell cardiac hypertrophy model using angiotensin Ⅱ (Ang Ⅱ ).Methods The cultured neonatal rat ventricular myocytes were treated with Ang Ⅱ as cell cardiac hypertrophy model.Hypertrophic neonatal rat cardiomyocytes were treated with GdCl3(a specific agonist of CaSR) and/or with Ro318220(a specific inhibitor of PKC pathway).To evaluate the status of cardiac hypertrophy,cell diameter was observed by HE dyeing,and protein content was determined through coomassie brilliant blue protein kit.The intracellular calcium concentration( [ Ca2+]i) was determined by laser scanning confocal microscope.The protein expression of CaSR and PKC pathway were analyzed using Western blotting.Results ①Compared to the control group(0.1263 ± 0.0443),the protein expression of CaSR was increased in Ang Ⅱ group and in GdCl3 group(0.1963 ± 0.0375,0.2778 ± 0.0564,all P＜ 0.05).Moreover,compared with Ang Ⅱ alone,the increase was significant in GdCl3 group(P ＜ 0.05).②Compared to control group(222.70 ± 22.09),AngⅡ group(392.16 ± 36.85) remarkably increased [Ca2+]i(P＜ 0.05),and this increase of [Ca2+]i was further enhanced in GdCl3 group (502.60 ± 44.21) versus Ang Ⅱ group (P ＜ 0.05).③Compared to control group,Ang Ⅱ could induce cardiomyocyte hypertrophy,and GdCl3 enhanced the effect.Moreover,this enhancement was attenuated by Ro318220.④Compared to control group(0.27 ± 0.07,0.69 ± 0.06,0.87 ± 0.04),the protein expression of PKCα,PKCε and PKCδ was increased in Ang Ⅱ group(0.60 ± 0.16,1.02 ± 0.13,1.20 ± 0.18,all P＜ 0.05) and the protein expression of PKCα,PKCε was increased in GdCl3 group(0.82 ± 0.16,1.34 ± 0.12,all P ＜ 0.05).Moreover,compared with Ang Ⅱ group,the protein expression of PKCα,PKCε was obviously increased in GdCl3 group (all P ＜ 0.05);compared with GdCl3 group,the protein expression of PKCα,PKCε(0.41 ± 0.10,0.85 ± 0.14) was obviously decreased in Ro318220 group(all P ＜ 0.05).Conclusions CaSR is involved in cardiac hypertrophy induced by Ang Ⅱ through PKC pathway in cultured neonatal rat cardiomyocytes.

Objective:To investigate the anti-oxidative stress effects of microRNA 125b (miR-125b) on lens epithelial cells (LECs) and its possible mechanism.Methods:Twenty-four anterior capsule specimens were collected from 24 eyes of 24 age-related cataract patients during phacoemulsification and 20 normal anterior capsule specimens were obtained from 20 eyes of 20 donors in Henan Eye Hospital from July 2018 to March 2019 under the approval of a Medical Ethics Committee of Henan Eye Hospital (No.YKYY20193151).The reverse transcription PCR and Western blot assay were employed to detect and compare the relative expression levels of miR-125b and nuclear factor E2-related factor 2 (Nrf2) in different specimens.The human lens epithelial cell line HLEB-3 was divided into control group and oxidative stress model group.The oxidative stress models were established by coculture with different concentrations (100, 200, 400 μmol/L) of H 2O 2 for 24 hours, and the cells were cultured with normal medium without H 2O 2 in the control group.The reactive oxygen species (ROS) content was detected by DCFH-DA fluorescent probe, and the activities of total-antioxidative capability (T-AOC), superoxide dismutase (SOD) and glutathione peroxidase (GSH-Px) as well as malondialdehyde (MDA) concentration were detected by ELISA, and compared among the groups.The expression levels of miR-125b and Nrf2 were detected by reverse transcription PCR and Western blot assay, respectively.The cells were transfected with miR-125b mimics, miR-125b control and miR-125b inhibitor for 24 hours, respectively, and ROS content was detected by DCFH-DA fluorescent probe and T-AOC, SOD and GSH-Px activities as well as MDA concentration were detected by ELISA and compared among different transfected groups.A dual luciferase reporter assay was used to assess an association between miR-125b and Nrf2.The expression level of Nrf2 protein was detected by Western blot assay and the expression levels of Nrf2 and Keap1 were assayed and located by immunofluorescence double staining. Results:The relative expression levels of miR-125b and Nrf2 in the normal lens anterior capsule specimens were 0.21±0.03 and 0.27±0.06, which were significantly lower than 0.89±0.05 and 0.84±0.12 in the cataract specimens, respectively ( t=15.355, P<0.05; t=18.647, P<0.05).The relative expression levels of miR-125b and Nrf2 were significantly increased in various H 2O 2 treated groups in comparison with the control group and were gradually elevated with the increase of H 2O 2 concentration (all at P<0.05).Compared with the control group, the T-AOC, SOD and GSH-Px activities were reduced, and ROS content and MDA concentration were significantly ascended (all at P<0.05).Compared with the miR-125b control group, the T-AOC, GSH-Px and SOD activities were increased, and ROS content and MDA concentration were decreased in the miR-125b mimics group (all at P<0.05).In addition, the T-AOC, GSH-Px and SOD activities were significantly weakened, and ROS content and MDA concentration were significantly increased in the miR-125b inhibitor group in comparison with the miR-125b control group (all at P<0.05).Dual luciferase reporter assay showed that miR-125b targeted to the expression of Nrf2 in the H 2O 2 model cells.The fluorescence of Nrf2 in the cytoplasm was the strongest with more nuclear transfer in the miR-125b mimics group, and the expression intensity of Keap1 in the cytoplasm was weaker.The expression of Nrf2 was the weakest with less nuclear transfer in the miR-125b inhibitor group, and the expression level of Keap1 in the cytoplasm was stronger. Conclusions:MiR-125b can enhance the anti-oxidative stress of LECs in age-related cataractous eyes probably by upregulating the expression of Nrf2 and activating the Keap1/Nrf2 signaling pathway.

The study of mammary gland bioreactor is in the ascendant. In order to generate transgenic goats of well-controlled expression of exogenic genes, we constructed a human lactoferrin (hLF) gene targeting vector containing promoter, exon 1, intron1 and some of exon 2 (about 6.1 kb fragment) and exon 6 approximately 9 (about 3.3 kb fragment) of the goat beta-casein gene as well as hLF minigene, neo gene inserted into them and tk gene ligated to the 3' end of the construct. The 9.4 kb goat genomic sequences as homologous arms were initially amplified by PCR with local goat tissue DNA. The expression vector was named pBC-tk-neo-hlf. Then the recombinant plasmid pBC-tk-neo-hlf containing hLF minigene was transfected into mice mammary tumor cell line C127 by liposome, cell clones were selected with G418. After proliferating, the transfected cells were induced with insulin, luteotropic hormone and hydrocortisone. The result of Western-blotting analysis showed that the transfected cells can secrete hLF protein, and the recombinant protein expressed in cultured cell supernatant has the similar molecular weight as the native protein. The expression level detected by ELISA was 0.21 microg/mL. This result indicated that the targeting vector could efficiently direct the expression of hLF in mammary cells,and it confirmed the validity of the constructed vector. At the same time, C127 cell line proved to be useful for evaluating the regulation of a foreign gene expression in mammary gland specific expression vector.
Animals ; Caseins ; genetics ; Cell Line, Tumor ; Goats ; Humans ; Lactoferrin ; genetics ; Mammary Glands, Animal ; cytology ; metabolism ; Mice ; Molecular Weight ; Transfection

OBJECTIVETo observe the effects of Xiaobai Decoction (XBD) in reducing albuminuria and shortening the duration of albuminuria in patients with pregnancy-induced hypertension syndrome (PIH) in puerperium.

METHODSEighty-five patients were given the conventional treatment with magnesium sulfate for relieving convulsion and lowering hypertension, at the same time, the treated group was given XBD additionally with the modification according to the symptoms. The treatment course for both groups was 14 days. Routine test of midstream urine was performed every three days, and 24 h-urinary protein was measured every week.

RESULTSThe therapeutic effect on the 43 patients of the treated group was markedly effective in 11 (25.6 % ), effective in 26 (60.4%) and ineffective in 6 cases (14.0%), the total effective rate being 86.0%; while in the 42 patients of the control group, the corresponding numbers were 5 (11.9%), 21 (50.0%), 16 (38.1%) and 61.9%, respectively, the efficacy of the former was significantly better (P < 0.05).

CONCLUSIONXBD is a simple, safe and effective drug for reducing albuminuria and shortening the duration of albuminuria in puerperium of PIH patients.

Adult ; Albuminuria ; drug therapy ; Drugs, Chinese Herbal ; therapeutic use ; Female ; Humans ; Hypertension, Pregnancy-Induced ; drug therapy ; Phytotherapy ; Postpartum Period ; Pregnancy ; Treatment Outcome ; Young Adult

OBJECTIVE: To evaluate the quality of reports published in recent 10 years in China about quantitative analysis of syndrome differentiation for diabetes mellitus (DM) in order to explore the methodological problems in these reports and find possible solutions. METHODS: The main medical literature databases in China were searched. Thirty-one articles were included and evaluated by the principles of clinical epidemiology. RESULTS: There were many mistakes and deficiencies in these articles, such as clinical trial designs, diagnosis criteria for DM, standards of syndrome differentiation of DM, case inclusive and exclusive criteria, sample size and estimation, data comparability and statistical methods. CONCLUSION: It is necessary and important to improve the quality of reports concerning quantitative analysis of syndrome differentiation of DM in light of the principles of clinical epidemiology.

OBJECTIVETo investigate the applications of percutaneous screw fixation for the treatment of pelvic fractures and its related surgical considerations.

METHODSFrom June 2010 to June 2012,19 patients with pelvic fractures were treated with percutaneous hollow screws. There were 13 males and 6 females, with an average age of 41 years (ranged from 22 to 58 years). Fractures were caused by traffic accidents in 11 cases, by falling down from high place in 8 cases. Based on the Tile classification, there were 15 cases of Tile C type and 4 case of Tile B type. The indexes such as screw inserting time, intraoperative blood loss, complications, functional recovery and reduction conditions were observed. Fixation methods included sacroiliac screws, cannulated screw fixation of the pubic ramus and cannulated screw fixation of the pubic symphysis separation.

RESULTSAnatomical reduction achieved in 7 cases, satisfactory reduction 11 cases, and unsatisfactory reduction 1 case. Union time of fracture union ranged from 8 to 12 weeks (mean 10 weeks). Wound infection,ununion of fracture and nerve injuries were not found. According to the Majeed standards, 12 patients obtained an excellent results, 6 good and 1 fair.

CONCLUSIONPercutaneous screw fixation for the treatment of pelvic fractures under fluoroscopy has several advantages such as less trauma, less blood loss, fewer rates of complications, reliable fixation and no blood transfusion, which can reconstruct the stability of the pelvic ring, but it needs adequate preoperative preparation and high requirements for the surgeon.

Adult ; Bone Screws ; Female ; Fracture Fixation, Internal ; Fractures, Bone ; diagnostic imaging ; surgery ; Humans ; Male ; Middle Aged ; Pelvic Bones ; diagnostic imaging ; injuries ; surgery ; Radiography ; Young Adult

Objective To study the inhibitory effects of TPP,a desmosdumotin-C B ring para-fluoro modified derivative,on human breast cancer MCF-7 cell proliferation,and investigate the possible mechanisms.Methods MTT assay was used to measure the proliferation suppression of MCF-7 cells after treated with 1.0,2.5,5.0,10.0,and 20.0 μg/mL TPP for 48 h,and then the cell apoptosis rate and expression rate of NF-κB P65 positive cells were tested by flow cytometry after 20.0 μg/mL TPP treatment for 0,24,and 48 h.Results MTT assay showed that,after treatment for 48 h,1.0,2.5,5.0,10.0,and 20.0 μg/mL TPP all exhibited the inhibitory effects and showed a dose-dependent relationship.Flow cytometry results showed that 20.0 μg/mL TPP induced cell apoptosis after treatment for 24 and 48 h.TPP (20.0 μg/mL) significantly reduced the rate ofNF-κB P65-positive cells in MCF-7 cells after treatment for 48 h.Conclusion TPP could inhibit the proliferation of MCF-7 cells,which may be induced by cell apoptosis.Down-regulation of NF-κB is possible to be related with apoptosis.